Anti-EMSY antibody (ab123)

Western blot - EMSY antibody (ab123)

WB using EMSY antibody (ab123) demonstrating the interaction between EMSY and BRCA2 by immunoprecipitation.

Endogenous coimmunoprecipitation of EMSY and BRCA2 from unmanipulated asynchronously dividing HeLa cells.

Immunoprecipitation with an unrelated antibody (anti-GFP), preimmune serum, ab123 or anti-BRCA2 antibody was followed by Western blotting with ab123.

Neither of the unrelated antibodies could coprecipitate EMSY, whereas an EMSY signal was detected in the immunoprecipitate obtained with the anti-BRCA2 antibody. The anti-EMSY antibody could also immunoprecipitate endogenous EMSY protein.

Hughes-Davies et al, 2003

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - EMSY antibody (ab123)

Image courtesy of Human Protein Atlas

ab123 staining EMSY in human fallopian tube tissue (nucleus of the glandular cells). Paraffin embedded tissue was incubated with ab123 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab123 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

Immunofluorescence - EMSY antibody (ab123)

Upper panel: EMSY responds to DNA damage.
EMSY antibody (ab123) immunofluorescence staining of immortalized mouse wild-type embryonic fibroblasts before and after treatment with ionizing radiation (10 Gray at 1.8 Gy/minute, 250 kVp). EMSY assembles into nuclear speckles over several hours.

Lower panel: Costaining with a monoclonal mouse antibody to gamma-H2AX reveals that EMSY relocalizes to DNA damage sites.

Hughes-Davies et al, 2003

Western blot

Western blot of EMSY on MCF-7 cell lysate.

Lane 1: ab4579 at 1/500
Lane 2: ab123 at 1/500.

Variability in the size at which EMSY runs by Western blot has been experienced with different protein marker systems. EMSY has been observed to run at between 115 and 150kD (even though the predicted size is 141kD). However, that the same size band is detected by both ab123 and ab4579 (raised with immunogens of different sequence) confirms the specificity of these antibodies.

Secondary ab Lane 1: Rabbit polyclonal to Goat IgG H&L (HRP) ab6741 (1/5000)
Exposure time: 3 min.

Secondary ab Lane 2: Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed ab7090 (1/5000)
Exposure time: 10 sec.

Lysate at 20µg/lane.

Lane 1 - Exposure time : 3 min.
Lane 2 - Exposure time : 10 sec.

Variability in the size at which EMSY runs by Western blot has been experienced with different protein marker systems. EMSY has been observed to run at between 115 and 150kD (even though the predicted size is 141kD). However, that the same size band is detected by both ab123 and ab4579 (raised with immunogens of different sequence) confirms the specificity of these antibodies.